靶向Hiwi基因的siRNA干扰对诱导三阴乳腺癌MDA-MB-231细胞凋亡的影响

The effect of inhibition of hiwi expression by RNAi on triple negative breast cancer cells MDA-MB-231 through inducing cell apoptosis

目的:探讨干扰Hiwi基因对三阴乳腺癌细胞MDA-MB-231凋亡的影响。方法:培养并转染三阴乳腺癌细胞MDA-MB-231,按照实验设计分为6组,转染后应用q RT-PCR、Western blot分析Hiwi基因的m RNA及其靶蛋白的表达情况,根据结果评估干扰效果筛选有效的si RNA干扰片段作为干扰组(Hiwi10330组),再按照试验设计分为3组分别是干扰组,阴性对照组(NC组)、空白对照组(Blank组),在细胞转染后进行细胞流式检测各实验组细胞凋亡率。结果:靶向Hiwi基因的si RNA干扰后,干扰组中m RNA的表达量明显降低(P<0.05),Hiwi基因靶蛋白的表达也出现了明显的抑制(P<0.05)。靶向Hiwi基因的si RNA干扰后,干扰组MDA-MB-231细胞的凋亡率与NC和Blank组相比明显升高(P<0.05)。结论:si RNA靶向沉默Hiwi基因后,乳腺癌细胞MDA-MB-231的凋亡率明显升高,提示Hiwi基因可能成为三阴乳腺癌的一潜在治疗靶点。

Objective To investigate the effect of interfering Hiwi gene on the apoptosis of MDA-MB-231 cells. Methods The mRNA and protein expression of Hiwi mRNA and its target protein were analyzed by qRT-PCR and Western Blot after transfection. MDA-MB- 231 cells were divided into 6 groups according to the experimental design. Interference effects were screened as siRNA interference group （Hiwi10330 group）, and then divided into 3 groups according to the experimental design： interference group, negative control group/NC, blank control group/ Blank. The cell apoptosis rate was detected by flow cytometry after transfection. Results The expression of mRNA in the interference group was significantly lower than that in the siRNA group （P 〈 0.05）, the expression of target protein of Hiwi gene was also significantly inhibited （P 〈 0.05）. The apoptosis rate of MDA- MB- 231 cells was significantly higher than that of NC and Blank groups （P 〈 0.05）. Conclusion The apoptosis rate of breast cancer cells MDA-MB-231 was significantly increased after siRNA targeting hiwi gene silencing.